The world is facing a major problem of antimicrobial resistance that is more severe in developing countries where the infectious diseases get spread more easily and the cost of treatment and prevention of them is very high. Multidrug resistance and extended spectrum β-lactamases (ESBL) production is a common problem, which needs empirical therapy to overcome and to reduce this drug resistance. In this study 88 clinical isolates were studied for ESBL production and presence of CTX-M genotypically. Out of 88 isolates, 32 were Escherichia coli (36%), 20 Klebsiella pneumoniae (23%), 14 Pseudomonas aeruginosa (16%), 12 Enterobacter cloacae (14%), 3 Proteus vulgaricus (3%), 5 Proteus mirabilis (7%) and 2 Acinetobacter baumannii (2%). Antibiotic susceptibility pattern showed 92% of the isolates were resistant to most beta-lactam antibiotics and plasmid was isolated from 72 isolates irrespective of its susceptibility pattern. 29 isolates were found to have CTX-M group I genes that includes 17 of 32 E. coli (53%), 8 of 20 K. pneumoniae (40%), 1 of 14 P. aeruginosa (7%) and 3 of 12 Enterobacter cloacae (25%). This study describes the genetic characteristics and molecular epidemiology of ESBLs among Enterobacteriaceae isolates and provides insight emergence of bacterial strains harbouring ESBL genes.